PROJECT SUMMARY ABSTRACT Despite therapeutic advances, neurodegenerative diseases and disorders remain some of the leading causes of mortality and morbidity in the United States. Therefore, cell-based therapies to replace lost or damaged neurons or supporting neural cells are of great therapeutic interest. Human neural progenitor cells (hNPCs) derived from human pluripotent stem cells (hPSCs, including human embryonic stem cells [hESCs] and human induced pluripotent stem cells [hiPSCs]) can proliferate extensively and differentiate into all the neural lineages (i.e. neurons, astrocytes, and oligodendrocytes) that compromise the central nervous system (CNS). Therefore, hNPCs and their differentiated progeny could provide the cellular raw material to model or treat a variety of nervous system disorders. However, the clinical application of these cells will require (i) defined, xeno-free conditions for their expansion and neuronal differentiation and (ii) scalable culture systems that enable their expansion and neuronal differentiation in numbers sufficient for regenerative medicine and drug screening purposes. To that end, we will use a novel high-throughput approach to systematically screen a rationally designed library of physicochemically-defined polymers to identify candidate synthetic substrates for the expansion and neuronal differentiation of hNPCs. Next, we will use these synthetic substrates as the basis for the engineering of low shear bioreactor-based systems for large-scale hNPC expansion and neuronal differentiation. The scaled-up cell populations will be assessed for their heterogeneity as well as their cellular, biochemical, genetic, and electrophysiological properties. The successful completion of this research will significantly advance the clinical application of hNPCs and their derivatives as it will enable the large-scale expansion and neuronal differentiation of hNPCs in quantities necessary for disease modeling, drug screening, and regenerative medicine applications.